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Background: Fibrosis is the most important predictor of prognosis in patients with Chronic hepatitis B (CHB) related liver disease. Detecting early fibrosis is vital to reduce complications of CHB like cirrhosis and HCC. High viral load is an important predictor of fibrosis. There are many studies reporting the association of hepatitis B (HBV) viral load with degree of liver fibrosis, but results are inconsistent. Hence, this study was undertaken to assess the relationship of HBV DNA titre with liver fibrosis. Methods: We recruited all newly detected treatment naive CHB patients between May 2021 to January 2023 at Department of Hepatology SCB medical college Cuttack. HBVDNA quantification and fibrosis assessment using 2D shear wave elastography (SWE) were done. The primary endpoint was the relation between HBVviral load and liver fibrosis stage and the estimation of HBV DNA cut off levels for advanced fibrosis ?F3. Results: Ninety-eight patients were enrolled (mean age: 46.05±13.21 years, male to female ratio-2.76:1). HBV DNA levels were higher in patients with advanced fibrosis LSM ?8kpa (F3) although the difference was statistically not significant (P=0.6). HBV DNA levels were also higher in CHB patients with significant fibrosis than those with less degree of fibrosis, in both HBeAg positive and HbeAg negative individuals, although this difference did not achieve statistical significance (p=0.655 and p=0.685). Age, serum albumin, serum ALT levels, total platelet count, CTP score and MELD-Na score were significant predictors of fibrosis on univariate analysis. But on multivariate analysis, platelet count (OR=0.999, CI=0.981-0.997, p=0.006) and MELD-Na score (OR=1.064, CI=1.01-1.198, p=0.024) were independent predictors of liver stiffness. A cut off HBV DNA of 17500 (IU/ML) with sensitivity of 51 % and specificity of 41% had a poor diagnostic accuracy to predict F3 fibrosis. Conclusion: There was no significant correlation between HBV DNA levels and stage of liver fibrosis.

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【Objective】 To explore the distribution of serological markers related to samples whose serological test results were inconsistent with HBV DNA test results among voluntary blood donors in Xi′an. 【Methods】 A total of 71 HBsAg ELISA positive and NAT non-reactive (ELISA+ /NAT-)blood samples were collected from Shaanxi Blood Center from November 1, 2022 to April 30, 2023. The serological markers of hepatitis B were detected by electrochemiluminescence method, and the HBV S region and C region gene fragments were amplified by nested-PCR. 【Results】 The positive rate of nested-PCR in double ELISA+ /NAT- group(n=30) was statistically higher than that of ELISA+ /NAT- group(n=41)(60% vs 24.4%, P<0.05). Donors in double ELISA+ /NAT- group were all first-time blood donors, with the positive rate of anti-HBc in serum of 100%, and the serological pattern was mainly positive for items 1, 4 and 5 items(80%). Among the ELISA+ /NAT- group, 31.7% were repeat blood donors, with the positive rate of anti-HBc in serum of only 19.51%, and the serological patterns were mainly single anti-HBs positive (43.90%) and all negative (36.58%). 【Conclusion】 There are false positives in the test results of ELISA+ /NAT- group, which leads to unnecessary blood discarding. Meanwhile, the samples with negative NAT may have low levels of HBV DNA, which may lead to missed detection. It is suggested that multiple systems and methods should be applied to trace the blood donors who are HBsAg positive and NAT non-reactive, so as to improve the accuracy of HBV screening of blood donors and reduce blood waste.

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【Objective】 To analyze the detection characteristics of a novel serum marker, hepatitis B core-associated antigen (HBcrAg), in the HBsAg-/HBV DNA+ blood donors in Wuxi. 【Methods】 A total of 37 previous HBsAg-/HBV DNA+ blood donors were followed up by telephone and their serum was obtained, and the serum of 22 HBsAg-/HBV DNA+ blood donors was detected by electrochemiluminescence and real-time PCR nucleic acid screening as the OBI group for HBcrAg enzyme-linked immunosorbent assay(ELISA). The serum of 20 healthy blood donors who underwent dual ELISA and one nucleic acid testing(NAT) was selected as the healthy control group, and the serum of 20 patients with chronic hepatitis B who were clinically diagnosed by Wuxi Fifth People's Hospital was selected as the experimental CHB group, and HBcrAg ELISA was detected respectively. The correlation analysis between HBcrAg and HBeAb, HBcAb, ALT and HBV DNA in the OBI group was performed. 【Results】 Thirty-seven blood samples were detected by chemiluminescence for HBsAg and NAT, and 22 HBsAg-/HBV DNA+ samples were detected in the OBI group, with a detection rate of 59.46%. The serum HBcrAg expression content (ng/mL) between the OBI group, the healthy control group and the CHB group were (0.92±0.13), (0.47±0.09) and (1.14±0.23), respectively, and the differences were statistically significant (P0.05). 【Conclusion】 The expression of HBcrAg in the OBI group and CHB group was higher than that in the healthy control group, and the serum HBcrAg was not correlated with HBeAb, HBcAb, ALT and HBV DNA to a certain extent. HBcrAg has a good application prospect in screening HBsAg-/HBV DNA+ blood donors.

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ObjectiveTo investigate the liver histopathological features of chronic hepatitis B （CHB） patients with normal alanine aminotransferase （ALT） and their correlation with serological markers. MethodsClinical data were collected from 137 patients with normal ALT who were treated in Wuxi Fifth People’s Hospital from April 2018 to June 2021， and the differences in liver histopathology and serological markers were analyzed， as well as the correlation between liver histopathology and serological markers. The chi-square test was used for comparison of categorical data between groups， and the Kruskal-Wallis H test was used for comparison of data between multiple groups. A Spearman rank correlation test was performed， and logistic regression was used to perform the multivariate analysis. ResultsIn the ALT ≤20 U/L， 20‍ ‍—‍ ‍29 U/L， and 30‍ ‍—‍ ‍40 U/L groups， the patients with significant inflammatory necrosis （≥G2） accounted for 57.4%， 53.4%， and 75%， respectively， and the patients with significant fibrosis （≥S2） accounted for 63.8%， 62.1%， and 75%， respectively. There was a significant difference in the degree of inflammatory necrosis between the patients with positive or negative HBeAg， the patients with different levels of serum HBV DNA， and the patients with different levels of serum HBV RNA （χ2=10.008， 6.911， and 7.946， all P<0.05）， and there was a significant difference in fibrosis stage between the patients with positive or negative HBeAg and the patients with different levels of serum HBV RNA （χ2=7.996 and 10.874， both P<0.05）. The degree of liver inflammation and fibrosis stage were not significantly correlated with serum HBV DNA （rs=0.024， P=0.785； rs=0.039， P=0.652）， while they were significantly correlated with serum HBV RNA （rs=0.222， P=0.009； rs=0.187， P=0.029）. The multivariate analysis showed that in CHB patients， positive HBeAg was an independent risk factor for inflammatory necrosis （odds ratio ［OR］=-0.302， 95% confidence interval ［CI］： -1.160 to 0.386， P=0.002） and fibrosis （OR=-0.387， 95%CI： -1.160 to 0.386， P=0.011）. ConclusionThere are varying degrees of inflammatory necrosis and fibrosis in the liver of CHB patients with normal ALT， and positive HBeAg is independent risk factor for significant inflammatory necrosis and fibrosis in liver tissue of these patients.

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@#Abstract: Objective　To investigate the correlation between HBV-DNA level, sterol O-acyltransferase (SOAT1) expression and tumor differentiation of hepatocellular carcinoma. Methods　The clinical and HBV-DNA level data from 58 cases of HBV-associated hepatocellular carcinoma were collected, and the cancer tissues and their paired paracancerous tissues were collected to detect SOAT1 expression by immunohistochemistry and evaluate tumor differentiation. Correlation was statistically analyzed using chi-square tests. Results　The high-level rate of HBV-DNA in the SOAT1 high expression group was 81.1% (30/37) compared to 19.1% (4/21) of the SOAT1 low expression group, with statistical significance, and there was also a correlation between SOAT1 expression and HBV-DNA levels (χ2=21.253,P<0.05). In the low differentiation hepatocellular carcinoma group, the rate of HBV-DNA high levels was 71.1% (27/38), while it was 35.0% (7/20) in the well-moderate differentiation group, with statistical significance. There was also a significant correlation between HBV-DNA levels and tumor differentiation degree (χ2=7.021,P<0.05). The overall positive rate of SOAT1 expression in all collected cases was 63.8% (37/58), with no expression (0/58) detected in all paired paracancerous tissues, with statistical significance (P<0.05). Furthermore, the expression level of SOAT1 protein in cancer tissues was correlated with the degree of tumor differentiation (χ2=19.889,P<0.05). SOAT1 was generally highly expressed in the low differentiated case group, with a positive rate of 84.2% (32/38), while SOAT1 was generally low expression or no expression in HCC samples with a higher degree of differentiation, with only a few samples exhibiting high expression, with a high expression rate of 25.0% (5/20). Conclusions　There is a correlation between HBV-DNA levels and hepatocellular carcinoma differentiation degree, with higher levels of HBV-DNA detected in low differentiation tumors. Additionally, the expression level of SOAT1 is also related to the degree of differentiation of hepatocellular carcinoma, and the expression level of SOAT1 in low differentiated carcinoma is also higher. Furthermore, there is a positive correlation between HBV-DNA levels and SOAT1 expression levels, and SOAT1 is a key enzyme involved in cellular lipid metabolism. These findings suggest that HBV infection may affect the function and level of SOAT1, which may interfere with hepatocyte lipid metabolism and participate in tumor genesis and evolution.

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【Objective】 To investigate the HBV infection markers detection and demographic characteristics of first-time blood donors, so as to provide evidence for blood donor recruitment. 【Methods】 HBsAg was detected by ELISA, HBsAg negative samples were tested for HBV DNA, and chemiluminescence method was used to detect HBsAg, HBsAb, HBcAb, HBeAg and HBeAb in first-time blood donor HBsAg-/HBV DNA+ samples. Demographic information of HBsAg positive first-time donors was analyzed. 【Results】 From 2018 to 2022, a total of 502 739 people participated in voluntary blood donation, and first-time blood donors accounted for 33.79%. The HBsAg positive rate of first-time donors(28.37/10 000, 482/169 897)was higher than that of repeated blood donors(3.46/10 000, 115/332 842) (OR=8.23, 95%CI: 6.72~10.09), and the HBV DNA positive rate of first-time blood donors(4.83/10 000, 82/169 897)was lower than that of repeated blood donors(6.52/10 000, 217/332 842)(OR=0.74, 95%CI: 0.57~0.95). The positive rate of HBcAb in HBsAg-/HBV DNA+ samples of first-time blood donors was 73.17%. Significant differences were noticed in HBsAg-/HBV DNA+ and HBsAg positive rate among first-time blood donors among gender, age, education background and occupation (all P<0.05). 【Conclusion】 Low risk first-time blood donor recruitment is important for blood donation. Strengthening HBV screening before blood donation and detection after blood donation is beneficial to improve the safety of blood transfusion.

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【Objective】 To analyze the hepatitis B virus (HBV) infection data of blood donors from 18 domestic blood stations, so as to investigate the HBV infection situation of blood donors. 【Methods】 The positive rate of HBV and its distribution characteristics of regions, the percentage of HBsAg+ ELISA in first-time vs repeated blood donors, and the percentage of HBsAg-/HBV DNA+ blood donors of 18 domestic blood stations during 2017 to 2020 were collected from the Working Platform for Practice Comparison of Blood Centers, and the HBV infection among blood donors were statistically analyzed. 【Results】 From 2017 to 2020, the positive rate of HBV in blood donors among 18 domestic blood stations was 13.48/10 000-144.02/10 000, with the average HBV positive rate in eastern, central and western region at 26.14/10 000, 51.98/10 000 and 41.00/10 000, respectively. The HBsAg+ rate by ELISA among first-time and repeated blood donors was 14.55/10 000-305.39/10 000 vs 1.04/10 000-87.43/10 000 The HBsAg-/HBV DNA+ yield was 1.80/10 000-35.31/10 000. 【Conclusion】 The distribution of HBV infection in blood donors has regional characteristics, and HBV prevalence was low in repeated blood donors. HBsAg ELISA combined with HBV DNA detection can better ensure blood safety.

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【Objective】 To detect and analyze the infection status of HBsAg non-reactive /HBV DNA reactive blood donors by individual donor-NAT (ID-NAT) and chemiluminescence technology, and to explore the feasibility and potential risks of reentry. 【Methods】 The blood screening results of blood donors in Wuhu from January 2018 to October 2021 were queried by blood station information management software. The blood donation information of all HBsAg non-reactive /HBV DNA reactive blood donors was collected and then recalled by telephone. After informed consent, samples were taken for HBV DNA nucleic acid single test, enzyme-linked immunoassay for HBsAg, chemiluminescence assay for HBV seromarkers(including HBsAg, anti-HBs, HBeAg, anti-HBe and anti-HBc), and alanine aminotransferase (ALT) test. All the results were statistically analyzed. 【Results】 From January 2018 to October 2021, there were 142 051 donations, and the positive rate of sole HBV DNA was 0.06% (91/142 051), and 33 people (37 person-times) were successfully followed up. The yield rates of HBsAg, anti-HBs and anti-HBc were 6.06% (2/33), 39.39% (13/33) and 96.97% (32/33), respectively; None HBeAg was yielded. After two times of ID-NAT, 8 patients remained non-reactive to both systems, with a negative conversion rate of 24.24% (8/33). Meanwhile, 25 patients were at least once reactive to ID-NAT, and 23 of them were occult HBV infection with serologically reactivity. There were 2(6.25%) patients with HBsAg positive conversion and HBV DNA persistent reactivity, which were window period infection. One person was confirmed as false reactivity (no HBV infection) as he remained unreactive to both repeated ID-NAT and serological tests. 【Conclusion】 Chemiluminescence assay is more sensitive than ELISA in detecting HBV serum markers, which is beneficial to early detection of HBV samples in window period. The yielding rate of anti-HBc among HBsAg non-reactive/HBV DNA reactive blood donors detected by blood screening in this region is very high, and most of them are occulting infection, so the ID-NAT should be no less than 2 times in the reentry strategy.

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ABSTRACT BACKGROUND: Occult hepatitis B virus infection (OBI) is defined as the presence of hepatitis B virus (HBV) deoxyribonucleic acid (DNA) in the liver of individuals with undetectable hepatitis B virus surface antigen (HBsAg) in the serum. The actual prevalence of OBI and its clinical relevance are not yet fully understood. OBJECTIVE: To evaluate the prevalence of HBV DNA in liver biopsies of HBsAg-negative patients with chronic liver disease of different etiologies in a referral center in Brazil and compare two different HBV DNA amplification protocols to detect HBV. DESIGN AND SETTING: This cross-sectional observational study was conducted at the Liver Outpatient Clinic, Hospital das Clínicas, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil, between January 2016 and December 2019. METHODS: HBV DNA was investigated in 104 liver biopsy samples from individuals with chronic liver disease of different etiologies, in whom HBsAg was undetectable in serum by nested-polymerase chain reaction (nested-PCR), using two different protocols. RESULTS: OBI, diagnosed by detecting HBV DNA using both protocols, was detected in 6.7% of the 104 individuals investigated. Both protocols showed a good reliability. CONCLUSION: In addition to the differences in the prevalence of HBV infection in different regions, variations in the polymerase chain reaction technique used for HBV DNA amplification may be responsible for the large variations in the prevalence of OBI identified in different studies. There is a need for better standardization of the diagnostic methods used to diagnose this entity.

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This article summarizes the risk of hepatocellular carcinoma in patients with chronic hepatitis B virus infection after HBsAg seroclearance, as well as its mechanism and implications.

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【Objective】 To analyze the status of HBV infection in blood donors reactive to jointed NAT but non-reactive to primary discriminatory tests (NRR), so as to provide suggestions and data support for subsequent studies on NRR samples. 【Methods】 HCV RNA and HIV RNA repeat differential detection, HBV DNA viral load detection and HBV pgRNA copy volume detection were performed in the plasma of 60 blood donors with negative ELISA results in routine blood screening and NRR in NAT. HBsAg, HBsAb, HBcAb, HBeAg and HBeAb serological tests were performed on the NRR samples with positivity in HBV DNA viral load and HBV pgRNA virus copy detection, so as to analyze the serological infection status and occult hepatitis B (OBI) infection. 【Results】 The HCV RNA and HIV RNA repeat discrimination results of 60 NRR samples were negative. The quantitative detection results of HBV DNA in 60 NRR samples were positive in 9 cases (15%), and the HBV DNA concentration was less than 10IU/mL. Nine cases (15%) were positive for HBV pgRNA quantitative detection, and the virus copy volume ±SD was (289±58.25) copies/mL. Two NRR samples (3.33%) were HBV DNA positive and HBV pgRNA positive. Among the 9 HBV-DNA positive samples, the highest positive rate of HBcAb was 66.67%, and 7 (77.78%) of them were confirmed to be seropositive for OBI. Among the 9 HBV pgRNA positive samples, the copy amount of pgRNA in HBcAb positive samples was slightly higher than that in negative samples, while the copy amount of pgRNA in HBsAb and HBeAb positive samples was lower than that in corresponding negative samples. In recent 6 years, the proportion of NRR samples in the single NAT system of the center fluctuated from 0.09% to 0.13%. 【Conclusion】 HBV DNA and HBV pgRNA exist in NRR samples. HBV DNA and/or HBV pgRNA positive samples can be detected in the relevant serological infectious markers. NRR samples have a certain potential risk of OBI infection. HBV DNA detection plus HBV pgRNA can better confirm the status of virus infection in NRR and improve the safety of blood transfusion.

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This study explored the correlation between interleukins (IL)-12, IL-18, and IL-21 and the viral load in patients with chronic hepatitis B virus (HBV). A total of 142 patients were consecutively enrolled. All were hepatitis B surface antigen (HBsAg)-positive for >6 months and did not receive drug therapy. An ELISA kit was used to test the IL-12, IL-18, IL-21, and acetylcholinesterase (AchE) levels in serum samples from chronic HBV patients and healthy control groups. The amounts of IL-12 and IL-18 were highest in the 5-6log10 (high viral load) group, while IL-21 was highest in the 3-4log10 (low viral load) group. Also, the IL-21 amount was decreased in the HBsAg+/HBeAg/HBcAb+ group, and IL-12, IL-18, and IL-21 were decreased in the normal alanine aminotransferase (ALT) group compared to the abnormal ALT group. These data suggested that IL-12, IL-18, and IL-21 serum levels were positively correlated with disease progression and could reflect disease severity for different HBV-DNA loads. Detection of IL-12, IL-18, and IL-21 levels was found to be helpful for evaluating the degree of liver cell damage and predicting the progression of hepatitis.

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【Objective】 To verify the results of HBV DNA and HCV RNA screening under different brands of vacuum collection tubes for blood samples, storage temperature and storage time. 【Methods】 Experiment 1 was conducted as follows: blood samples were collected simultaneously from 52 voluntary blood donors using two brands(divided into group A and group B) of vacuum collection tubes for blood samples. The plasma separation of group A and group B were compared, and the effects of storage time on the NAT yield of HBV DNA and HCV RNA were statistically analyzed. Experiment 2 was conducted as follows: the effects of different storage temperature, time and tubes on the NAT yield of HBV DNA and HCV RNA samples with low viral load in group A and B were verified and compared in the simulated phlebotomy condition. 【Results】 In Experiment 1: After centrifugation, blood plasma layer and cells layer were separated completely in group A(100%, 52/52), but one sample was not well separated in group B(1/52, 1.92%). After 4 to 10 h after collection, blood samples of two groups were centrifuged and screened for HBV DNA, HCV RNA within 24 h. No positive samples were yielded and the Ct values of internal control(IC-DNA and IC-RNA) were uniform. In Experiment 2: Whole blood samples, stored for either 4 h or 6~10 h at 4 ℃ or 25℃ before centrifugation, showed no difference on the NAT-yield of HBV DNA nor HCV RNA samples with low viral load(P>0.05). Ct values of HBV DNA and HCV RNA of group A was similar to those of group B as centrifuged samples were stored for 24 h or 72~104 h at 4℃(P>0.05), but all increased as the storage time prolonged. Ct values of HBV DNA in group A increased from 33.45±0.29(24 h) to 33.82±0.08(72~104 h) and HCV RNA from 35.21±0.20 to 36.12±0.43; HBV DNA from 33.46±0.25 to 34.30±0.60 and HCV RNA from 35.47±0.24 to 36.49±0.51 in group B. 【Conclusion】 Under certain laboratory condition, different storage time, storage temperature and tubes shed few effect on the NAT-yield of HBV DNA and HCV RNA samples with low virus loads. However, it is suggested that the blood sample be detected within 72 h after centrifugation at 4 ℃ storage.

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【Objective】 To compare the quantitative detection results of two domestic quantitative real-time PCR reagents in HBV-DNA detection. 【Methods】 A total of 306 serum samples form hepatitis B patients were selected for quantitative parallel detection of high-sensitiveity HBV-DNA using domestic reagent A and B, and the difference and consistency of the results were analyzed. 【Results】 1) The yielding rate of reagent A and B was 86.92% and 84.64%, respectively. The regression linear equation was Y=0.984 9x+ 0.154 9, R2=0.945 7, the correlation coefficient r=0.972 5, indicating the results by the two reagents had good correlation. 2) When the concentration was in the range of(20~1 000) IU/mL, the yielding rates of reagent A and B were 39.9% and 37.6%, respectively. 【Conclusion】 Reagent A is more suitable for post-treatment monitoring in patients with OBI and HBeAg(-), but also can be used to detect HBV pathogens in patients before operations or blood transfusions.

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Introducción: Los ensayos para cuantificar el ADN del virus de la hepatitis B (VHB) o carga viral son imprescindibles en el diagnóstico y en el seguimiento de los pacientes con hepatitis B crónica; de ahí que estén disponibles estuches diagnósticos para esta función. En el presente estudio se muestra la validación de SUMASIGNAL VHB (un paso), el cual es un sistema de reacción en cadena de la polimerasa en tiempo real (RCP-TR) para la cuantificación del genoma del VHB, propuesto por el Centro de Inmunoensayo. Objetivo: Evaluar el desempeño analítico de SUMASIGNAL VHB (un paso). Métodos: Se utilizó un panel de 80 muestras de suero bien caracterizadas y el Tercer Estándar Internacional de la OMS para las técnicas de amplificación de ácidos nucleicos del virus de la hepatitis B. Se determinaron las características del ensayo como especificidad clínica, especificidad analítica (reactividad cruzada), rango lineal o linealidad y exactitud, precisión intraensayo y comparación con un ensayo de referencia. Resultados: La especificidad analítica y clínica fue del 100 por ciento. Al evaluar la linealidad y exactitud con un estándar de referencia de la OMS, se obtuvo que la totalidad de las diferencias entre los Log10 del valor obtenido y el de referencia resultaron inferiores a 0,5 Log10 (r= 0,9977 y r2= 0,9954). Además, se obtuvieron bajos coeficientes de variación intraensayo. La evaluación comparativa con el estuche comercial Artus HBV RG PCR kit mostró una correlación fuerte (r= 0,8882). Conclusiones: SUMASIGNAL VHB (un paso) es un ensayo fácil de realizar manualmente, es rápido e incluye reactivos de extracción de ácidos nucleicos. Teniendo en cuenta la validez del método para el uso previsto, puede ser recomendado para su introducción en el diagnóstico, la vigilancia y la indicación de tratamiento en los pacientes con hepatitis B crónica(AU)

Introduction: Assays to quantify hepatitis B virus (HBV) DNA or viral load are indispensable for the diagnosis and follow-up of patients with chronic hepatitis B, hence the availability of diagnostic kits for this purpose. The present study deals with the validation of HBV SUMASIGNAL (one step), a real time polymerase chain reaction (RT-PCR) system for quantification of the HBV genome proposed by the Immunoassay Center. Objective: Evaluate the analytical performance of HBV SUMASIGNAL (one step). Methods: Use was made of a panel of 80 well characterized serum samples and the Third WHO International Standard for hepatitis B virus nucleic acid amplification techniques. Determination was performed of assay characteristics such as clinical specificity, analytical specificity (cross-reactivity), linear range or linearity and accuracy, intra-assay precision and comparison with a reference assay. Results: Analytical and clinical specificity was 100 percent. Evaluation of linearity and accuracy with a WHO reference standard revealed that all the differences between the log10 of the value obtained and the reference value were lower than 0.5 log10 (r= 0.9977 and r2= 0.9954). The intra-assay variation coefficients obtained were low. Comparative evaluation with the commercial Artus HBV RG PCR kit showed a strong correlation (r= 0.8882). Conclusions: The assay HBV SUMASIGNAL (one step) is easy to conduct manually, fast and includes reagents for nucleic acid extraction. Based on the validity of the method for the use in mind, it may be recommended for incorporation into the diagnosis, surveillance and treatment of patients with chronic hepatitis B(AU)

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ObjectiveTo investigate the clinical significance of highly sensitive nucleic acid detection in precise antiviral therapy for patients with liver cirrhosis and its association with aminotransferase level. MethodsA total of 377 patients with hepatitis B cirrhosis who were hospitalized or attended the outpatient service from May 2013 to April 2019 were enrolled and tested by both domestic HBV DNA detection and highly sensitive Cobas HBV DNA detection. All patients underwent biochemical examination, four blood coagulation tests, routine blood test, and upper abdominal computed tomography or ultrasound. Sensitivity of different HBV DNA detection reagents was compared in liver cirrhosis patients with a low viral load, and the correlation between alanine aminotransferase (ALT) level and viral load was analyzed. The paired t-test was used for comparison of continuous data before and after treatment. The receiver operating characteristic (ROC) curve was used to screen out the optimal predictive values of ALT at different cut-off values of HBV DNA. ResultsAmong the 377 patients with hepatitis B cirrhosis, 215 tested positive and 162 tested negative by domestic HBV DNA, and among these 162 patients, 104 (64.2%) tested positive by Cobas HBV DNA detection, with a mean level of 267.5±42.3 IU/ml. After 24 weeks of antiviral therapy, the 104 patients with hepatitis B cirrhosis had significant improvements in viral replication level, ALT, and Child-Pugh score for liver function; HBV DNA decreased from 267.5±32.2 IU/ml before treatment to 59.6±7.7 IU/ml after treatment (t=3.486, P=0.002), ALT decreased from 871±10.8 U/L before treatment to 36.5±7.6 U/L after treatment (t=3.235, P=0.020), and the Child-Pugh score decreased from 6.5±0.7 before treatment to 5.7±0.5 after treatment (t=2.928, P=0.041). The ROC curve analysis of ALT in predicting HBV DNA decision point showed that an ALT level of 29 IU/L was the most sensitive cut-off value for predicting HBV DNA ＜20 IU/ml, with an area under the ROC curve of 0.904, a sensitivity of 1.0, and a specificity of 0.237. ConclusionPrecise detection helps to guarantee the precise clinical treatment of patients with hepatitis B cirrhosis and improve their treatment outcome and prognosis. An ALT level of 29 IU/L is a sensitive indicator for predicting patients with negative Cobas HBV DNA, so as to achieve individualized precise screening and treatment.

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The detection and quantification of hepatitis B virus (HBV) DNA plays an important role in diagnosing and monitoring HBV infection as well as in assessing the therapeutic response. We compared the analytical performance of a random access, fully automated HBV assay—DxN VERIS Molecular Diagnostics System (Beckman Coulter, Brea, CA, USA)—with that of Abbott RealTime HBV assay (Abbott Laboratories, Des Plaines, IL, USA). The between-day precision of the VERIS assay ranged from 0.92% (mean 4.68 log IU/mL) to 4.15% (mean 2.09 log IU/mL) for pooled sera from HBV patients. HBV DNA levels measured by the VERIS HBV assay correlated with the calculated HBV DNA levels (r²=0.9994; P < 0.0001). The lower limit of quantification was estimated as 8.76 IU/mL (Probit analysis, 95% confidence interval: 7.32–12.00 IU/mL). Passing-Bablok regression analysis showed good concordance between the VERIS and RealTime assays for 187 chronic HBV samples (y=−0.2397+0.9712x; r=0.981), as well as for 20 drug-resistant HBV genotype C positive samples (y=−0.5415+0.9954x; r=0.961). The VERIS assay demonstrated performance similar to the RealTime assay and is suitable for high-throughput HBV DNA monitoring in large hospital laboratories.

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Objective To reveal the characteristics of S gene sequence of hepatitis B surface antigen (HBsAg) in hepatitis B virus (HBV)-infected patients with low HBsAg level.Methods From February 2016 to December 2017, 1 308 serum samples of inactive HBsAg carriers were collected from the 903rd Hospital of PLA and Hangzhou Jianggan District People′s Hospital.The cases were divided into high-level group and low-level group according to the level of serum HBsAg (10 IU／mL) expression.The HBV S gene was sequenced in patients with low-level HBsAg expression.In addition, in patients with high-level HBsAg, 100 patients were randomly selected (stratified sampling) for HBV S gene sequencing based on the matching of age and serological pattern (hepatitis B e antigen [HBeAg] negative) of low-level HBsAg group.A comparative analysis was conducted between HBV S gene sequences from inactive HBsAg carrier in low HBsAg expression group and the HBV reference S gene sequences from inactive HBsAg carrier in high HBsAg expression group .The results of normal distribution data were expressed as Mean ±SD, and analyzed using t-test.The results of non-normal distribution data were expressed by M(QR), and analyzed using Mann-Whitney U test.Chi-square test or Fisher exact test was used to compare continuous variables and classification variables between the two groups .Results There were 276 serum samples from the low level group and 1 032 serum samples from the high level group , including 257 HBsAg／HBeAg／anti-HBc-positive cases, 753 HBsAg／anti-HBe／anti-HBc-positive cases, and 22 HBsAg／anti-HBc-positive cases.Successful HBV S gene sequencing was performed on 126 out of 276 patients in the low-level HBsAg group.According to the age inthe low-level HBsAg group, 100 samples with negative HBeAg in the high-level HBsAg group were randomly selected , among which 94 patients were genotyped and hemotyped.The results showed that there were statistically significant differences in HBV serological markers , HBV DNA level and HBV genotype distribution between the high level group (94 cases) and the low level group (126 cases) (all P＜0.05).The ASC-R-B and ASC-R-C genotypes reported in this study had high homology (99.6%-100.0%) with those reported in Shanghai , Chengdu, Wuhan, Yunnan and Beijing of China , and high homology (98.2%-99.6%) with those reported in Japan and Korea of NCBI genotype B and C reference sequences, but had low homology with patients far away from China (98.2% in Canada and 98.7% in Indonesia).In genotype B of the low level group , the amino acid mutation number of SHB protein was 71, and the hot spot mutation number was 19, both higher than those in the high level group (39 and 8, respectively). The difference was statistically significant (χ2 ＝12.303 and 4.766, respectively, both P＜0.05).Amino acid mutation sites in the low HBsAg group were mainly distributed on both sides of the major hydrophilic region (MHR) (amino acid residues 40 -49 and 198 -220).There were no significant differences in amino acid mutation number and hot spot mutation number between the two groups of C genotype (χ2 ＝0.383 and 0.409, respectively, both P＞0.05).For genotype B, 12 single point mutations and 4 dual co-mutations were found in low level group.Among them, one single point mutation (S210R) and 3 dual co-mutations (G44E／V+T45P／I, G44E／V+L49P／R and N40S+I208T) were not hot spot mutations , while 2 dual co-mutations and 2 single point mutations were found in high level group.The difference between two groups was statistical significant (χ2 ＝7.533,P ＝0.006).For genotype C, 5 single point mutations ( T5A, A45T, T47A／K, Q101R and I126S／T) were found in low level group and 1 single point mutation (N3S) in high level group.The difference in mutation frequency between two groups were statistical significant (χ2 ＝47.914,P＝0.000).Conclusions Significant mutations in multiple regions and at multiple sites ( including co-mutations) on both sides of the MHR may be one of the causes of low HBsAg expression level in this population .

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Introduction@#Worldwide, an estimated two billion people have evidence of HBV infection, and approximately 240 million have CHB. In this study, a representative group of Mongolian adults was tested for hepatitis B virus (HBV) in 2017. The prevalence estimates of HBV the general Mongolian adult population were found to be 11.1%, respectively.</br> In April 2017, EASL added a drug newly approved for treatment of CHB, tenofovir alafenamide (TAF) to their list of recommended first-line therapies. The requirement for long-term therapy in chronic HBV highlights the importance of these efficacy and safety trends, however their true clinical relevance is yet to be established and further studies with long-term follow up and real-world clinical data are needed.@*Goal@#Evaluate for result of tenofovir alafenamide in the treatment of chronic hepatitis B infection.@*Materials and Methods@#The clinical trials have evaluated TAF in HBeAg-positive and HBeAg-negative chronic HBV patients. The trials have similar designs and are randomized, double blind, non-inferiority studies. The primary efficacy endpoint was the proportion of patients with HBV DNA<29 IU/ml at week 24 and 48. Other prespecified efficacy endpoints were the proportion of patients with HBsAg seroncoversion to anti-HBs at week 24 and 48. Key secondary safety end- points at week 24 and 48 included the percentage change in T-score, and Z-score bone mineral density (BMD), percentage change in BMD and change from baseline serum creatinine.@*Results@#The primary efficacy endpoint, an HBV DNA level <29 IU/ml at week 24, was achieved by 120 (59.1%) of 203 patients receiving TAF, which was non-inferior to the 63 (55.2%) of 114 patients receiving TDF who had an HBV DNA<29 IU/ml. After 24 weeks of treatment, patients receiving TAF had significantly smaller reductions in bone mineral density (BMD) compared with patients receiving TDF.@*Conclusion@#The development of TAF, specifically designed to deliver potent antiviral activity but with an improved safety profile compared with TDF, is therefore timely.

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@#Worldwide, an estimated two billion people have evidence of HBV infection, and approximately 240 million have CHB. In April 2017, EASL added a drug newly approved for treatment of CHB, tenofoviralafenamide (TAF) to their list of recommended first-line therapies. Treatment with these therapies can achieve sustained suppression of HBV DNA replication, decreases in inflammation, and histological activity that decrease the risk of cirrhosis and hepatocellular carcinoma in both cirrhotic and noncirrhotic patients and, ultimately, of CHB-associated mortality [1, 2]. However, recent advances in understanding the HBV life cycle have enabled multiple, novel therapeutic targets to be identified and new therapies of direct-acting antiviral (DAAs) and host-targeting agents (HTAs) are indevelopment.</br> In most clinical trials, TAF was non-inferior to TDF in achieving HBV DNA levels below 29 IU/ml.No amino-acid substitutions associated with viral breakthrough were detected by deep sequencing, and no resistance to TAF.With clear evidence from major studies showing that TAF is safe, tolerable, and non-inferior to TDF, its recommendation as a first-line therapy is appropriate.</br> Long-term safety is an important consideration in the therapeutic management of patients with CHB because treatment is often life-long.</br> The efficacy of TAF in patients with resistance mutations associated with older nucleos(t)ide analogues is unclear. Although no evidence of TAF or TDF resistance was detected in the phase III studies through 96 weeks of treatment, very small numbers of patients had baseline mutations indicating resistance to lamivudine, adefovir or entecavir and efficacy data specifically for this group is not available.